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United States Patent | 6,103,521 |
Capon ,   et al. | August 15, 2000 |
Novel multispecific chimeric receptor DNA sequences, expression cassettes and vectors containing these sequences as well as cells containing the chimeric DNA and novel chimeric receptor proteins expressed from the sequences are provided in the present invention. The novel multispecific chimeric receptor DNA and amino acid sequences comprise at least three domains that do not naturally exist together: (1) a multispecific binding domain comprising at least two extracellular inducer-responsive clustering domains which serves to bind at least one specific inducer molecule, (2) a transmembrane domain, which crosses the plasma membrane, and (3) either a proliferation signaling domain that signals the cell to divide, or an effector function signaling domain which directs a host cell to perform its specialized function. Optionally, all the multispecific chimeric receptors may contain one or more intracellular inducer-responsive clustering domains attached to one or more of the cytoplasmic signaling domains or the transmembrane domain. The present invention also relates to novel hybrid multispecific chimeric receptors comprising at least one proliferation signaling domain and at least one effector function signaling domain together on the multispecific receptor molecule. The present invention further relates to therapeutic methods and strategies that employ the cells expressing these novel chimeric receptors for the treatment of cancer, infectious disease and autoimmune disease which may have greater therapeutic benefit over a combination of drug therapies.
Inventors: | Capon; Daniel J. (Hillsborough, CA); Smith; Douglas H. (Foster City, CA); Tian; Huan (Cupertino, CA); Winslow; Genine A. (Hayward, CA); Siekevitz; Miriam (New York, NY) |
Assignee: | Cell Genesys, Inc. (Foster City, CA) |
Appl. No.: | 454098 |
Filed: | May 30, 1995 |
Current U.S. Class: | 435/325; 435/69.7; 435/320.1; 530/350; 530/387.3; 536/23.4 |
Intern'l Class: | C12N 015/62; C07K 014/705 |
Field of Search: | 536/23.4 435/69.1,240.2,252.3,320.1,325,69.7 530/350,351,387.1,387.3 |
5260203 | Nov., 1993 | Ladner et al. | 435/172. |
5336603 | Aug., 1994 | Capon et al. | 435/69. |
5359046 | Oct., 1994 | Capon et al. | 536/23. |
5741899 | Apr., 1998 | Capon et al. | 536/23. |
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TABLE 1 ______________________________________ Construct Concentration (.mu.g/ml) ______________________________________ CD4-Fc 1.6 SAb(.alpha.gp41)-Fc 1.2 SAb(.alpha.gp120-Fc 1.3 SAb(.alpha.gp41)-SAb(.alpha.gp120)-Fc 0.23 SAb(.alpha.gp41)-L1-SAb(.alpha.gp120)-Fc 0.65 SAb(.alpha.gp41)-L2-SAb(.alpha.gp120)-Fc 0.75 SAb(.alpha.gp41)-L3-SAb(.alpha.gp120)-Fc 0.40 SAb(.alpha.gp41)-L4-SAb(.alpha.gp120)-Fc 0.48 SAb(.alpha.gp120)-CD4-Fc 0.05 SAb(.alpha.gp120)-L1-CD4-Fc 0.17 SAb(.alpha.gp120)-L2-CD4-Fc 0.19 SAb(.alpha.gp120)-L3-CD4-Fc 0.10 SAb(.alpha.gp120)-L4-CD4-Fc 0.12 ______________________________________
__________________________________________________________________________ # SEQUENCE LISTING - - - - (1) GENERAL INFORMATION: - - (iii) NUMBER OF SEQUENCES: 26 - - - - (2) INFORMATION FOR SEQ ID NO:1: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 36 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1: - - CTGCGTCAAC ACAGACTGTG AGGAGACGGT GACCAG - # - # 36 - - - - (2) INFORMATION FOR SEQ ID NO:2: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: - - ACAGACTGTG AGGAGA - # - # - # 16 - - - - (2) INFORMATION FOR SEQ ID NO:3: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 78 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: - - CTGCGTCAAC ACAGACTGAC CCTTACCCTC AGAAGATTTA CCCGACCCCG AG - #GTCGACCC 60 - - TGAGGAGACG GTGACCAG - # - # - # 78 - - - - (2) INFORMATION FOR SEQ ID NO:4: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: - - AGAAGATTTA CCCGAC - # - # - # 16 - - - - (2) INFORMATION FOR SEQ ID NO:5: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 111 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: - - CTGCGTCAAC ACAGACTGAC CGTCCTTCTT AGCGTCGTCC TTCTTAGCGT CG - #TCCTTCTT 60 - - AGCAGCGTCC TTCTTAGCGT CGTCAGCGGA AGATGAGGAG ACGGTGACCA G - # 111 - - - - (2) INFORMATION FOR SEQ ID NO:6: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: - - GCGTCGTCCT TCTTAG - # - # - # 16 - - - - (2) INFORMATION FOR SEQ ID NO:7: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 66 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: - - CTGCGTCAAC ACAGACTGTG GGGACGGTGG GGATGTGTGA GTTTTGTCTG AG - #GAGACGGT 60 - - GACCAG - # - # - # 66 - - - - (2) INFORMATION FOR SEQ ID NO:8: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: - - CGGTGGGGAT GTGTGA - # - # - # 16 - - - - (2) INFORMATION FOR SEQ ID NO:9: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 87 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: - - CTGCGTCAAC ACAGACTGGT CCAGCTCCCC GTCCTGCGCT TCGGCGCTCG AT - #TCTTCCAG 60 - - TTGCAGCTCT GAGGAGACGG TGACCAG - # - # 87 - - - - (2) INFORMATION FOR SEQ ID NO:10: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10: - - TCGGCGCTCG ATTCTT - # - # - # 16 - - - - (2) INFORMATION FOR SEQ ID NO:11: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 36 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11: - - GCCCAGCACC ACTTTCTTTG AGCTCACGGT GACCGT - # - # 36 - - - - (2) INFORMATION FOR SEQ ID NO:12: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: - - ACTTTCTTTG AGCTCA - # - # - # 16 - - - - (2) INFORMATION FOR SEQ ID NO:13: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 78 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: - - GCCCAGCACC ACTTTCTTAC CCTTACCCTC AGAAGATTTA CCCGACCCCG AG - #GTCGACCC 60 - - TGAGCTCACG GTGACCGT - # - # - # 78 - - - - (2) INFORMATION FOR SEQ ID NO:14: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 111 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: - - GCCCAGCACC ACTTTCTTAC CGTCCTTCTT AGCGTCGTCC TTCTTAGCGT CG - #TCCTTCTT 60 - - AGCAGCGTCC TTCTTAGCGT CGTCAGCGGA AGATGAGCTC ACGGTGACCG T - # 111 - - - - (2) INFORMATION FOR SEQ ID NO:15: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 66 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15: - - GCCCAGCACC ACTTTCTTTG GGGACGGTGG GGATGTGTGA GTTTTGTCTG AG - #CTCACGGT 60 - - GACCGT - # - # - # 66 - - - - (2) INFORMATION FOR SEQ ID NO:16: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 87 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: - - GCCCAGCACC ACTTTCTTGT CCAGCTCCCC GTCCTGCGCT TCGGCGCTCG AT - #TCTTCCAG 60 - - TTGCAGCTCT GAGCTCACGG TGACCGT - # - # 87 - - - - (2) INFORMATION FOR SEQ ID NO:17: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 42 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: - - TAGTCTAGGA TCTACTGGCT GCAGTTCTTG CTCTCCTCTG TC - # - # 42 - - - - (2) INFORMATION FOR SEQ ID NO:18: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: - - ACTGGCTGCA GTTCTT - # - # - # 16 - - - - (2) INFORMATION FOR SEQ ID NO:19: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 39 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: - - AAAACTGTGC GTTACAATTC GTGGGTCCCC TCCTGAGGA - # - # 39 - - - - (2) INFORMATION FOR SEQ ID NO:20: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20: - - TACAATTCGT GGGTCC - # - # - # 16 - - - - (2) INFORMATION FOR SEQ ID NO:21: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 39 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21: - - TCCTATTGTA ACAAATGCTT GCCCTGGTCC TCTCTGGAT - # - # 39 - - - - (2) INFORMATION FOR SEQ ID NO:22: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base - #pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: DNA (genomic) - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22: - - AAATGCTTGC CCTGGT - # - # - # 16 - - - - (2) INFORMATION FOR SEQ ID NO:23: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 amino - #acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: peptide - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: - - Gly Ser Thr Ser Gly Ser Gly Lys Ser Ser Gl - #u Gly Lys Gly 1 5 - # 10 - - - - (2) INFORMATION FOR SEQ ID NO:24: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 25 amino - #acids (B) TYPE: amino acid (C) STRANDEDNESS: single
(D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: peptide - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24: - - Ser Ser Ala Asp Asp Ala Lys Lys Asp Ala Al - #a Lys Lys Asp Asp Ala 1 5 - # 10 - # 15 - - Lys Lys Asp Asp Ala Lys Lys Asp Gly 20 - # 25 - - - - (2) INFORMATION FOR SEQ ID NO:25: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 amino - #acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: peptide - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25: - - Asp Lys Thr His Thr Ser Pro Pro Ser Pro 1 5 - # 10 - - - - (2) INFORMATION FOR SEQ ID NO:26: - - (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 amino - #acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear - - (ii) MOLECULE TYPE: peptide - - (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26: - - Glu Leu Gln Leu Glu Glu Ser Ser Ala Glu Al - #a Gln Asp Gly Glu Leu 1 5 - # 10 - # 15 - - Asp __________________________________________________________________________